Production and characterization of a highly pure RNA polymerase holoenzyme from Mycobacterium tuberculosis
Descripción del Articulo
Recent publications have shown that active RNA polymerase (RNAP) from Mycobacterium tuberculosis (MtbRNAP) can be produced by expressing all four subunits in a single recombinant Escherichia coli strain [1–3]. By reducing the number of plasmids and changing the codon usage of the Mtb genes in the co...
| Autores: | , , , , , |
|---|---|
| Formato: | artículo |
| Fecha de Publicación: | 2017 |
| Institución: | Consejo Nacional de Ciencia Tecnología e Innovación |
| Repositorio: | CONCYTEC-Institucional |
| Lenguaje: | inglés |
| OAI Identifier: | oai:repositorio.concytec.gob.pe:20.500.12390/751 |
| Enlace del recurso: | https://hdl.handle.net/20.500.12390/751 https://doi.org/10.1016/j.pep.2017.03.013 |
| Nivel de acceso: | acceso abierto |
| Materia: | recombinant protein bacterial protein DNA directed RNA polymerase holoenzyme biosynthesis chemistry enzymology Escherichia coli genetic transcription https://purl.org/pe-repo/ocde/ford#3.00.00 |
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CONC_65f2b7a8ac1630475afc5dcebd64f993 |
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oai:repositorio.concytec.gob.pe:20.500.12390/751 |
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CONC |
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CONCYTEC-Institucional |
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4689 |
| dc.title.none.fl_str_mv |
Production and characterization of a highly pure RNA polymerase holoenzyme from Mycobacterium tuberculosis |
| title |
Production and characterization of a highly pure RNA polymerase holoenzyme from Mycobacterium tuberculosis |
| spellingShingle |
Production and characterization of a highly pure RNA polymerase holoenzyme from Mycobacterium tuberculosis Herrera-Asmat O. recombinant protein bacterial protein DNA directed RNA polymerase holoenzyme biosynthesis chemistry enzymology Escherichia coli genetic transcription https://purl.org/pe-repo/ocde/ford#3.00.00 |
| title_short |
Production and characterization of a highly pure RNA polymerase holoenzyme from Mycobacterium tuberculosis |
| title_full |
Production and characterization of a highly pure RNA polymerase holoenzyme from Mycobacterium tuberculosis |
| title_fullStr |
Production and characterization of a highly pure RNA polymerase holoenzyme from Mycobacterium tuberculosis |
| title_full_unstemmed |
Production and characterization of a highly pure RNA polymerase holoenzyme from Mycobacterium tuberculosis |
| title_sort |
Production and characterization of a highly pure RNA polymerase holoenzyme from Mycobacterium tuberculosis |
| author |
Herrera-Asmat O. |
| author_facet |
Herrera-Asmat O. Lubkowska L. Kashlev M. Bustamante C.J. Guerra D.G. Kireeva M.L. |
| author_role |
author |
| author2 |
Lubkowska L. Kashlev M. Bustamante C.J. Guerra D.G. Kireeva M.L. |
| author2_role |
author author author author author |
| dc.contributor.author.fl_str_mv |
Herrera-Asmat O. Lubkowska L. Kashlev M. Bustamante C.J. Guerra D.G. Kireeva M.L. |
| dc.subject.none.fl_str_mv |
recombinant protein |
| topic |
recombinant protein bacterial protein DNA directed RNA polymerase holoenzyme biosynthesis chemistry enzymology Escherichia coli genetic transcription https://purl.org/pe-repo/ocde/ford#3.00.00 |
| dc.subject.es_PE.fl_str_mv |
bacterial protein DNA directed RNA polymerase holoenzyme biosynthesis chemistry enzymology Escherichia coli genetic transcription |
| dc.subject.ocde.none.fl_str_mv |
https://purl.org/pe-repo/ocde/ford#3.00.00 |
| description |
Recent publications have shown that active RNA polymerase (RNAP) from Mycobacterium tuberculosis (MtbRNAP) can be produced by expressing all four subunits in a single recombinant Escherichia coli strain [1–3]. By reducing the number of plasmids and changing the codon usage of the Mtb genes in the co-expression system published by Banerjee et al. [1], we present a simplified, detailed and reproducible protocol for the purification of recombinant MtbRNAP containing the ω subunit. Moreover, we describe the formation of ternary elongation complexes (TECs) with a short fluorescence-labeled RNA primer and DNA oligonucleotides, suitable for transcription elongation studies. |
| publishDate |
2017 |
| dc.date.accessioned.none.fl_str_mv |
2024-05-30T23:13:38Z |
| dc.date.available.none.fl_str_mv |
2024-05-30T23:13:38Z |
| dc.date.issued.fl_str_mv |
2017 |
| dc.type.none.fl_str_mv |
info:eu-repo/semantics/article |
| format |
article |
| dc.identifier.uri.none.fl_str_mv |
https://hdl.handle.net/20.500.12390/751 |
| dc.identifier.doi.none.fl_str_mv |
https://doi.org/10.1016/j.pep.2017.03.013 |
| dc.identifier.scopus.none.fl_str_mv |
2-s2.0-85016006342 |
| url |
https://hdl.handle.net/20.500.12390/751 https://doi.org/10.1016/j.pep.2017.03.013 |
| identifier_str_mv |
2-s2.0-85016006342 |
| dc.language.iso.none.fl_str_mv |
eng |
| language |
eng |
| dc.relation.ispartof.none.fl_str_mv |
Protein Expression and Purification |
| dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess |
| eu_rights_str_mv |
openAccess |
| dc.publisher.none.fl_str_mv |
Elsevier B.V. |
| publisher.none.fl_str_mv |
Elsevier B.V. |
| dc.source.none.fl_str_mv |
reponame:CONCYTEC-Institucional instname:Consejo Nacional de Ciencia Tecnología e Innovación instacron:CONCYTEC |
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Consejo Nacional de Ciencia Tecnología e Innovación |
| instacron_str |
CONCYTEC |
| institution |
CONCYTEC |
| reponame_str |
CONCYTEC-Institucional |
| collection |
CONCYTEC-Institucional |
| repository.name.fl_str_mv |
Repositorio Institucional CONCYTEC |
| repository.mail.fl_str_mv |
repositorio@concytec.gob.pe |
| _version_ |
1844883116666126336 |
| spelling |
Publicationrp01914600rp01915600rp01916600rp01918600rp01316500rp01917600Herrera-Asmat O.Lubkowska L.Kashlev M.Bustamante C.J.Guerra D.G.Kireeva M.L.2024-05-30T23:13:38Z2024-05-30T23:13:38Z2017https://hdl.handle.net/20.500.12390/751https://doi.org/10.1016/j.pep.2017.03.0132-s2.0-85016006342Recent publications have shown that active RNA polymerase (RNAP) from Mycobacterium tuberculosis (MtbRNAP) can be produced by expressing all four subunits in a single recombinant Escherichia coli strain [1–3]. By reducing the number of plasmids and changing the codon usage of the Mtb genes in the co-expression system published by Banerjee et al. [1], we present a simplified, detailed and reproducible protocol for the purification of recombinant MtbRNAP containing the ω subunit. Moreover, we describe the formation of ternary elongation complexes (TECs) with a short fluorescence-labeled RNA primer and DNA oligonucleotides, suitable for transcription elongation studies.Consejo Nacional de Ciencia, Tecnología e Innovación Tecnológica - ConcytecengElsevier B.V.Protein Expression and Purificationinfo:eu-repo/semantics/openAccessrecombinant proteinbacterial protein-1DNA directed RNA polymerase-1holoenzyme-1biosynthesis-1chemistry-1enzymology-1Escherichia coli-1genetic transcription-1https://purl.org/pe-repo/ocde/ford#3.00.00-1Production and characterization of a highly pure RNA polymerase holoenzyme from Mycobacterium tuberculosisinfo:eu-repo/semantics/articlereponame:CONCYTEC-Institucionalinstname:Consejo Nacional de Ciencia Tecnología e Innovacióninstacron:CONCYTEC#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#20.500.12390/751oai:repositorio.concytec.gob.pe:20.500.12390/7512024-05-30 15:44:23.699http://purl.org/coar/access_right/c_14cbinfo:eu-repo/semantics/closedAccessmetadata only accesshttps://repositorio.concytec.gob.peRepositorio Institucional CONCYTECrepositorio@concytec.gob.pe#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#<Publication xmlns="https://www.openaire.eu/cerif-profile/1.1/" id="69188c2d-6e0e-406a-96c9-34bb92b2f3ba"> <Type xmlns="https://www.openaire.eu/cerif-profile/vocab/COAR_Publication_Types">http://purl.org/coar/resource_type/c_1843</Type> <Language>eng</Language> <Title>Production and characterization of a highly pure RNA polymerase holoenzyme from Mycobacterium tuberculosis</Title> <PublishedIn> <Publication> <Title>Protein Expression and Purification</Title> </Publication> </PublishedIn> <PublicationDate>2017</PublicationDate> <DOI>https://doi.org/10.1016/j.pep.2017.03.013</DOI> <SCP-Number>2-s2.0-85016006342</SCP-Number> <Authors> <Author> <DisplayName>Herrera-Asmat O.</DisplayName> <Person id="rp01914" /> <Affiliation> <OrgUnit> </OrgUnit> </Affiliation> </Author> <Author> <DisplayName>Lubkowska L.</DisplayName> <Person id="rp01915" /> <Affiliation> <OrgUnit> </OrgUnit> </Affiliation> </Author> <Author> <DisplayName>Kashlev M.</DisplayName> <Person id="rp01916" /> <Affiliation> <OrgUnit> </OrgUnit> </Affiliation> </Author> <Author> <DisplayName>Bustamante C.J.</DisplayName> <Person id="rp01918" /> <Affiliation> <OrgUnit> </OrgUnit> </Affiliation> </Author> <Author> <DisplayName>Guerra D.G.</DisplayName> <Person id="rp01316" /> <Affiliation> <OrgUnit> </OrgUnit> </Affiliation> </Author> <Author> <DisplayName>Kireeva M.L.</DisplayName> <Person id="rp01917" /> <Affiliation> <OrgUnit> </OrgUnit> </Affiliation> </Author> </Authors> <Editors> </Editors> <Publishers> <Publisher> <DisplayName>Elsevier B.V.</DisplayName> <OrgUnit /> </Publisher> </Publishers> <Keyword>recombinant protein</Keyword> <Keyword>bacterial protein</Keyword> <Keyword>DNA directed RNA polymerase</Keyword> <Keyword>holoenzyme</Keyword> <Keyword>biosynthesis</Keyword> <Keyword>chemistry</Keyword> <Keyword>enzymology</Keyword> <Keyword>Escherichia coli</Keyword> <Keyword>genetic transcription</Keyword> <Abstract>Recent publications have shown that active RNA polymerase (RNAP) from Mycobacterium tuberculosis (MtbRNAP) can be produced by expressing all four subunits in a single recombinant Escherichia coli strain [1–3]. By reducing the number of plasmids and changing the codon usage of the Mtb genes in the co-expression system published by Banerjee et al. [1], we present a simplified, detailed and reproducible protocol for the purification of recombinant MtbRNAP containing the ω subunit. Moreover, we describe the formation of ternary elongation complexes (TECs) with a short fluorescence-labeled RNA primer and DNA oligonucleotides, suitable for transcription elongation studies.</Abstract> <Access xmlns="http://purl.org/coar/access_right" > </Access> </Publication> -1 |
| score |
13.4721 |
Nota importante:
La información contenida en este registro es de entera responsabilidad de la institución que gestiona el repositorio institucional donde esta contenido este documento o set de datos. El CONCYTEC no se hace responsable por los contenidos (publicaciones y/o datos) accesibles a través del Repositorio Nacional Digital de Ciencia, Tecnología e Innovación de Acceso Abierto (ALICIA).
La información contenida en este registro es de entera responsabilidad de la institución que gestiona el repositorio institucional donde esta contenido este documento o set de datos. El CONCYTEC no se hace responsable por los contenidos (publicaciones y/o datos) accesibles a través del Repositorio Nacional Digital de Ciencia, Tecnología e Innovación de Acceso Abierto (ALICIA).
